Orthogonal Crosslinking for Protein Stabilization and Cytosolic Delivery

用于蛋白质稳定和胞质递送的正交交联

• 批准号: 1904558
• 负责人: Qing Lin
• 金额: $ 42.6万
• 依托单位: SUNY at Buffalo
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-15 至 2023-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1904558&HistoricalAwards=false
• 关键词: Orthogonal; Crosslinking; Protein; Stabilization; Cytosolic

Large quantities of proteins can readily be produced from bacteria and, therefore, represent a sustainable source of biological catalysts and therapeutics. With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Qing Lin from the State University of New York at Buffalo to develop a chemical modification to proteins that, when used with light, increases their stability and their ability to enter cells. This chemical biology strategy enables the design of protein-based catalysts with increased resistance to structural disruptions from heat and chemicals. In addition, this new modification enables the development of cell-penetrating small proteins derived from antibodies (called nanobodies) for the diagnosis and treatment of disease. The project provides interdisciplinary research opportunities to undergraduate students, including some from the groups underrepresented in STEM fields, through collaborations with the New York State Collegiate Science and Technology Entry Program.Site-specific incorporation of a 2-aryl-5-carboxytetrazole (ACT) linked amino acid into a protein allows the creation of head-to-tail circular proteins through intramolecular photo-crosslinking with a proximal nucleophilic residue. ACT-based orthogonal inter-strand crosslinking is also explored in this project for the synthesis of nanobodies that display positively charged surfaces to enhance cell-penetration. The crosslinking efficiency and selectivity of the various constructs are optimized through chemical synthesis. The effects of orthogonal crosslinking on protein conformational stability and cytosolic uptake are assessed using appropriate biological assays. These studies are expected to yield new insights into the reactivity of photo-generated reactive species inside proteins, which leads to designed proteins with novel structures and functions.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

细菌可以很容易地产生大量蛋白质，因此是生物催化剂和治疗药物的可持续来源。凭借该奖项，化学部的生命过程化学项目正在资助纽约州立大学布法罗分校的 Qing Lin 博士开发一种对蛋白质的化学修饰，当与光一起使用时，可以提高其稳定性和进入细胞的能力。这种化学生物学策略使得基于蛋白质的催化剂的设计能够增强对热和化学物质引起的结构破坏的抵抗力。此外，这种新的修饰使得能够开发源自抗体（称为纳米抗体）的细胞穿透小蛋白，用于疾病的诊断和治疗。该项目通过与纽约州大学科学技术入门计划合作，为本科生提供跨学科研究机会，包括一些来自 STEM 领域代表性不足的群体的学生。将 2-芳基-5-羧基四唑 (ACT) 连接的氨基酸定点掺入蛋白质中，可以通过分子内光交联创建头尾环状蛋白质。 近端亲核残基。该项目还探索了基于 ACT 的正交链间交联，用于合成具有带正电表面的纳米抗体，以增强细胞渗透。通过化学合成优化各种构建体的交联效率和选择性。使用适当的生物测定法评估正交交联对蛋白质构象稳定性和胞质摄取的影响。这些研究预计将对蛋白质内部光生反应物种的反应性产生新的见解，从而设计出具有新颖结构和功能的蛋白质。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Orthogonal Crosslinking: A Strategy to Generate Novel Protein Topology and Function.

正交交联：生成新型蛋白质拓扑和功能的策略。

• DOI: 10.1002/chem.202202828
• 发表时间: 2023
• 期刊: Chemistry (Weinheim an der Bergstrasse, Germany)
• 作者: Wang,Zheng;Rabb,JohnathanD;Lin,Qing
• 通讯作者: Lin,Qing

Spontaneous Orthogonal Protein Crosslinking via a Genetically Encoded 2-Carboxy-4-Aryl-1,2,3-Triazole.

• DOI: 10.1002/anie.202202657
• 发表时间: 2022-05-23
• 期刊: ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
• 影响因子: 16.6
• 作者: Xu, Yali;Rahim, Abdur;Lin, Qing
• 通讯作者: Lin, Qing

Light-Triggered Click Chemistry.

• DOI: 10.1021/acs.chemrev.0c00799
• 发表时间: 2021-06-23
• 期刊: Chemical reviews
• 影响因子: 62.1
• 作者: Kumar GS;Lin Q
• 通讯作者: Lin Q

Fitness Factors for Bioorthogonal Chemical Probes.

• DOI: 10.1021/acschembio.9b00755
• 发表时间: 2019-12-20
• 期刊: ACS chemical biology
• 影响因子: 4
• 作者: Tian Y;Lin Q
• 通讯作者: Lin Q